Screening neuraminidase inhibitors from glycosaminoglycan and natural extract by capillary electrophoresis

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• 作者：Ting-Fu Jiang ; Lei Chong ; Mei-E Yue ; Yuan-Hong Wang…
• 关键词：neuraminidase inhibitor ; screening ; capillary electrophoresis ; electrophoretically mediated microanalysis
• 刊名：Journal of Analytical Chemistry
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：71
• 期：3
• 页码：283-288
• 全文大小：526 KB
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• 作者单位：Ting-Fu Jiang (1) (2)
  Lei Chong (1) (2)
  Mei-E Yue (3)
  Yuan-Hong Wang (1) (2)
  Zhi-Hua Lv (1) (2)
  
  1. School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, P.R. China
  2. Key Laboratory of Marine Drugs, Ministry of Education of China, Qingdao, 266003, P.R. China
  3. College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, 266042, P.R. China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Russian Library of Science
  
• 出版者：MAIK Nauka/Interperiodica distributed exclusively by Springer Science+Business Media LLC.
• ISSN：1608-3199

文摘

An electrophoretically mediated microanalysis (EMMA) method for screening neuraminidase inhibitors in depolymerized glycosaminoglycan and natural extracts is described. In the present method, enzyme and substrate were individually introduced into the capillary as distinct plugs, and then mixed for a short time. Afterwards the voltage was reapplied to separate the product from the unreacted substrate and the natural extract. The measured peak area of the product at 214 nm represents the enzyme activity. The electrophoretic conditions for the enzyme reaction and separation of substrate and product were optimized in this study. Under the optimal conditions, the Michaelis–Menten constant and the inhibitive mechanism of zanamivir were studied, which agreed with the literature data. Furthermore, the inhibitory ratios of enzymatic activity of depolymerized glycosaminoglycan and traditional Chinese drugs were determined. The EMMA method has superiority over traditional assay methods, in not only minimizing the false-positive results but also in simplifying the experimental procedure. Therefore, it could be employed to screen inhibitors from natural sources.